Evidence map›Paper›PMID 25460726›Full record

ArticleRedox biology2014

Protein engineering to develop a redox insensitive endothelial nitric oxide synthase.

Ruslan Rafikov, Sanjiv Kumar, Saurabh Aggarwal, Daniel Pardo, Fabio V Fonseca, Jessica Ransom, Olga Rafikova, Qiumei Chen, Matthew L Springer, Stephen M Black

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Redox biology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed.

  1. Article
  2. Article
  3. Article
  4. Hyper-activation of pp60Free radical biology & medicine · 2017
    Article
  5. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

10 authors.

Ruslan RafikovPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Sanjiv KumarPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Saurabh AggarwalPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Daniel PardoPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Fabio V FonsecaPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Jessica RansomPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Olga RafikovaPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA.
Qiumei ChenThe Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, USA.
Matthew L SpringerThe Division of Cardiology, University of California, San Francisco, San Francisco, CA, USA.
Stephen M BlackPulmonary Vascular Disease, Vascular Biology Center, Georgia Regents University, Augusta, GA, USA. Electronic address: sblack@gru.edu.

Funding

Therapeutic Mechanisms of RhoA Inhibition In Acute Lung InjuryP01HL101902 · NHLBI · AUGUSTA UNIVERSITY · PI BLACK, STEPHEN M · 2011 to 2016
$11.2M
PERINATAL REGULATION OF THE ENDOTHELIAL NOS GENER01HL060190 · NHLBI · UNIVERSITY OF MONTANA · PI Stephen M Black · 1998 to 2026
$7.1M
ROS in Pulmonary Hypertension: Role of ADMAR01HL067841 · NHLBI · UNIVERSITY OF MONTANA · PI BLACK, STEPHEN M · 2002 to 2011
$2.6M
The Role of Nitric Oxide Synthase in Circulating Progenitor Cell FunctionR01HL086917 · NHLBI · UNIVERSITY OF CALIFORNIA, SAN FRANCISCO · PI SPRINGER, MATTHEW LAWRENCE · 2007 to 2011
$2.1M
Perinatal regulation of TGF-beta1 during vascular remodelingR01HL084739 · NHLBI · AUGUSTA UNIVERSITY · PI BLACK, STEPHEN M · 2007 to 2010
$1.5M
Role of altered carnitine metabolism in perinatal endothelial dysfunctionR21HD057406 · NICHD · AUGUSTA UNIVERSITY · PI BLACK, STEPHEN M · 2008 to 2009
$423k
Endothelin and EGFR Activation in Pulmonary HypertensionF32HL103136 · NHLBI · AUGUSTA UNIVERSITY · PI RAFIKOVA, OLGA · 2010 to 2012
$115k
NHLBI NIH HHS F32 HL103136NHLBI NIH HHS P01 HL101902NHLBI NIH HHS R01 HL060190NHLBI NIH HHS R01 HL067841NHLBI NIH HHS R01 HL084739NHLBI NIH HHS R01 HL086917NICHD NIH HHS R21 HD057406
6 · The paper itself

Abstract

The zinc tetrathiolate (ZnS4) cluster is an important structural feature of endothelial nitric oxide synthase (eNOS). The cluster is located on the dimeric interface and four cysteine residues (C94 and C99 from two adjacent subunits) form a cluster with a Zn ion in the center of a tetrahedral configuration. Due to its high sensitivity to oxidants this cluster is responsible for eNOS dimer destabilization during periods of redox stress. In this work we utilized site directed mutagenesis to replace the redox sensitive cysteine residues in the ZnS4 cluster with redox stable tetra-arginines. Our data indicate that this C94R/C99R eNOS mutant is active. In addition, this mutant protein is insensitive to dimer disruption and inhibition when challenged with hydrogen peroxide (H2O2). Further, the overexpression of the C94R/C99R mutant preserved the angiogenic response in endothelial cells challenged with H2O2. The over-expression of the C94R/C99R mutant preserved the ability of endothelial cells to migrate towards vascular endothelial growth factor (VEGF) and preserved the endothelial monolayer in a scratch wound assay. We propose that this dimer stable eNOS mutant could be utilized in the treatment of diseases in which there is eNOS dysfunction due to high levels of oxidative stress.

Indexed as

Amino Acid SubstitutionMutation, MissenseNitric Oxide Synthase Type IIIAnimalsChlorocebus aethiopsCOS CellsHumansHydrogen PeroxideHydrogen PeroxideNitric Oxide Synthase Type IIINOS3 protein, humanEndothelial nitric oxide synthaseProtein engineeringRedox stabilityZinc tetrathiolate cluster

Identifiers

PMID25460726
PMCPMC4297941

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.